An Italian Multicenter Perspective Harmonization Trial for the Assessment of MET Exon 14 Skipping Mutations in Standard Reference Samples

• Published in: Diagnostics (Basel) 2023-02, Vol.13 (4), p.629
• Main Authors: Bironzo, Paolo, Pepe, Francesco, Russo, Gianluca, Pisapia, Pasquale, Gragnano, Gianluca, Aquino, Gabriella, Bessi, Silvia, Buglioni, Simonetta, Bartoccini, Federico, Ferrero, Giuseppina, Bresciani, Michela Anna, Francia di Celle, Paola, Sibona, Francesca, Giusti, Andrea, Movilia, Alessandra, Farioli, Renata Mariella, Santoro, Alessandra, Salemi, Domenico, Scarpino, Stefania, Galafate, Dino, Tommasi, Stefania, Lacalamita, Rosanna, Seminati, Davide, Sajjadi, Elham, Novello, Silvia, Pagni, Fabio, Troncone, Giancarlo, Malapelle, Umberto
• Format: Article
• Language: English
• Subjects: Acids; Automation; Biomarkers; Causes of; Data analysis; Diagnosis; Exon (Molecular genetics); Health aspects; Kinases; Laboratories; Lung cancer; MET; molecular test; Mutation; Mutation (Biology); NSCLC; Pathology; Polymerase chain reaction; Proprietary; Signal transduction; Software
• ISSN: 2075-4418; 2075-4418
• DOI: 10.3390/diagnostics13040629

Lung cancer remains the leading cause of cancer deaths worldwide. International societies have promoted the molecular analysis of MET proto-oncogene, receptor tyrosine kinase ( ) exon 14 skipping for the clinical stratification of non-small cell lung cancer (NSCLC) patients. Different technical approaches are available to detect exon 14 skipping in routine practice. Here, the technical performance and reproducibility of testing strategies for exon 14 skipping carried out in various centers were evaluated. In this retrospective study, each institution received a set ( = 10) of a customized artificial formalin-fixed paraffin-embedded (FFPE) cell line (Custom ex14 skipping FFPE block) that harbored the exon 14 skipping mutation (Seracare Life Sciences, Milford, MA, USA), which was previously validated by the Predictive Molecular Pathology Laboratory at the University of Naples Federico II. Each participating institution managed the reference slides according to their internal routine workflow. exon 14 skipping was successfully detected by all participating institutions. Molecular analysis highlighted a median Cq cut off of 29.3 (ranging from 27.1 to 30.7) and 2514 (ranging from 160 to 7526) read counts for real-time polymerase chain reaction (RT-PCR) and NGS-based analyses, respectively. Artificial reference slides were a valid tool to harmonize technical workflows in the evaluation of exon 14 skipping molecular alterations in routine practice.